The invention relates generally to the field of nucleic acid synthesis and, more specifically, to methods and systems for synthesizing nucleic acids and quality control of such synthesis.
Recombinant nucleic acid techniques have facilitated the study of isolated genes in a wide range of organisms. For example, such techniques have allowed researchers to express a particular gene (e.g., a mouse gene) in a host organism (e.g., bacteria) to study not only the effects of the gene itself, but also to study its expression products. While such techniques have been used to study naturally-occurring nucleic acids, researchers have also been interested in studying the effects of particular mutations or changes in a nucleic acid sequence. For example, targeted mutations may be introduced into a nucleic acid sequence that in turn result in a change in an expressed protein sequence. Researchers may then study the effects of such mutations on protein interactions of interest. Such mutations may be introduced through PCR-based techniques and the resulting sequences may be expressed in host cells. However, these techniques may involve labor-intensive analysis and time-consuming host cell propagation to determine if a particular host cell has incorporated the sequence of interest. Further, while recombinant techniques may be suited for simply generating mutations (e.g. point mutations, deletions, or substitutions) of a pre-existing nucleic acid sequence, such techniques are not well suited to creating an entirely synthetic, template-independent nucleic acid molecule.
Some nucleic acid synthesis techniques are chemically-based or enzymatic techniques, e.g., chemical solid phase synthesis, that involve adding individual nucleotides to one another in the desired order to form a polynucleotide chain, such as a PCR primer of limited length (e.g., 30-50 nucleotides). However, nucleic acid synthesis techniques suitable for forming relatively short sequences may be slow and/or error-prone when used for synthesis of a long polynucleotide chain (e.g., greater than 50 nucleotides). For example, techniques with an error-rate of 1/500 nucleotides may yield generally error-free short sequences of under 50 nucleotides, but may introduce several errors when used to synthesize sequences of 500-2000 nucleotides.